DOTAP chloride
(Synonyms: (2,3-二油氧基丙基)三甲基氯化铵,1,2-Dioleoyl-3-trimethylammoniumpropane (chloride)) 目录号 : GC41811
DOTAP chloride 是一种脂质体制剂，已用于将质粒 DNA 转染到培养的真核细胞中 [1]。
Cas No.:132172-61-3
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
mammalian CV-1 and 3T3 cells |
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Preparation Method |
Lipid dispersions containing cationic lipid analogues were prepared by first bath-sonicating dried lipid samples in distilled water to clarity (2-5 min), then adding an equal concentration of 308 mM NaCl, 40 mM Hepes (pH 7.4), and sonicating further for 2 min. Lipid/DNA mixtures for transfections were prepared by incubating dotap chloride with DNA for 5 min, typically at a total lipid concentration of 0.5-2 mg/ml, before addition to cell monolayers. Reverse-phase evaporation vesicles were prepared as described previously [2] and filtered through 0.1/xm pore-size Nucleopore filters. Lipid mixing between cationic lipid dispersions and large unilamellar PC/PS vesicles was assayed by a resonance energy transfer-based procedure as described previously [3], incubating cationic lipid dispersions (2 μM), colabeled with 1 mol% (12-CPS)-18 PC and 0.35 mol% (12-DABS)-18 PC, with a 9-fold excess (18 μM) of unlabeled PC/PS vesicles. For transfection of cells using cationic lipid dispersions, cell monolayers were washed twice with HEPESbuffered saline, then incubated with lipid/DNA mixtures in the latter medium (total volume 3 ml per100-mm culture dish) at 37 °C, for 90 min except whereotherwise indicated. Following this incubation period,the incubation medium was either replaced or supplemented with 12 ml of D-MEM containing 5% serum,and the cells were cultured for a further 36 h beforeharvesting. Chloramphenicol acetyltransferase activityin plasmid pSV2cat-transfected cells was assayed asdescribed previously [4], using chloramphenicol andacetyl-CoA concentrations of 3.5 μM and 2.5 mM,respectively. |
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Reaction Conditions |
0.5-2 mg/ml for 90 min |
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Applications |
For transfection of the cells with a fixed amount of plasmid (usually 2 or 5 µg) |
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References: [1]: Leventis R, Silvius J R. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles[J]. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1990, 1023(1): 124-132. [2]: Wilschut J, Duzgunes N, Fraley R, et al. Studies on the mechanism of membrane fusion: kinetics of calcium ion induced fusion of phosphatidylserine vesicles followed by a new assay for mixing of aqueous vesicle contents[J]. Biochemistry, 1980, 19(26): 6011-6021. [3]: Silvius J R, Leventis R, Brown P M, et al. Novel fluorescent phospholipids for assays of lipid mixing between membranes[J]. Biochemistry, 1987, 26(14): 4279-4287. [4]: Gorman C M, Moffat L F, Howard B H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells[J]. Molecular and cellular biology, 1982, 2(9): 1044-1051. |
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DOTAP chloride is a liposome formulation that have been used to transfect plasmid DNA into eukaryotic cells in culture [1]. The synthetic cationic amphiphile DOTAP chloride has been shown to mediate uptake of plasmid also in the presence of a culture medium containing serum. It is metabolized in the cells and has proven to be less cytotoxic than other amphiphiles [2]. DOTAP chloride has also been used successfully for internalization of an 18-mer DNA oligonucleotide, and it was shown in these experiments to protect the oligonucleotide from degradation by nucleases in serum and cells [3].
References:
[1]. Haukenes G, Szilvay A M, Brokstad K A, et al. Labeling of RNA transcripts of eukaryotic cells in culture with BrUTP using a liposome transfection reagent (DOTAP?)[J]. Biotechniques, 1997, 22(2): 308-312.
[2]. Leventis R, Silvius J R. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles[J]. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1990, 1023(1): 124-132.
[3]. Capaccioli S, Dipasquale G, Mini E, et al. Cationic lipids improve antisense oligonucleotide uptake and prevent degradation in cultured cells and in human serum[J]. Biochemical and biophysical research communications, 1993, 197(2): 818-825.
DOTAP chloride 是一种脂质体制剂，已用于将质粒 DNA 转染到培养的真核细胞中 [1]。合成的阳离子两亲物 DOTAP chloride 已被证明在含有血清的培养基存在下也能介导质粒的摄取。它在细胞内代谢，已证明其细胞毒性低于其他两亲物 [2]。 DOTAP 氯化物也已成功用于 18 聚体 DNA 寡核苷酸的内化，并且在这些实验中表明它可以保护寡核苷酸免于被血清和细胞中的核酸酶降解[3]。
| Cas No. | 132172-61-3 | SDF | |
| 别名 | (2,3-二油氧基丙基)三甲基氯化铵,1,2-Dioleoyl-3-trimethylammoniumpropane (chloride) | ||
| Canonical SMILES | CCCCCCCC/C=C\CCCCCCCC(OC(C[N+](C)(C)C)COC(CCCCCCC/C=C\CCCCCCCC)=O)=O.[Cl-] | ||
| 分子式 | C42H80NO4•Cl | 分子量 | 698.6 |
| 溶解度 | ≤33mg/ml in ethanol;0.5mg/ml in DMSO;5mg/ml in dimethyl formamide | 储存条件 | Store at -20°C, protect from light, stored under nitrogen |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.4314 mL | 7.1572 mL | 14.3143 mL |
| 5 mM | 0.2863 mL | 1.4314 mL | 2.8629 mL |
| 10 mM | 0.1431 mL | 0.7157 mL | 1.4314 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Non-viral dried powders for respiratory gene delivery prepared by cationic and chitosan loaded liposomes
Int J Pharm2008 Nov 19;364(1):108-18.PMID: 18775770DOI: 10.1016/j.ijpharm.2008.07.034
The aim of this work was to investigate lipid-based dried powders as transfection competent carriers capable of promoting the expression of therapeutic genes. The lipid-based vectors were prepared by combining different cationic lipids 1,2-dioleoyl-3-trimethylammoniumpropane chloride (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 3beta(N(N',N-dimethylaminoethane) carbamoyl) cholesterol hydrochloride (DC-Chol) or by mixing of anionic lipids (1,2-dimyristoyl-sn-glycero-3-phospocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phospho-rac-glycerol sodium salt (DMPG) and chitosan salts. Spray drying of the formulations was performed using carbohydrates as thermoprotectant excipients and some amino acids as aerosolisation enhancers. Both the lipidic vectors and the dried powders were characterized for morphology, size, zeta potential (Z-potential) and a yield of the process. Agarose gel electrophoresis was used to examine the structural integrity of dehydrated plasmid DNA (pDNA). The biological functionality of the powders was quantified using the in vitro cell transfection. Among the several lipids and lipid-polymer mixtures tested, the best-selected formulations had spherical shape, narrow size distribution (mean diameter<220 nm, P.I.<0.250), a positive zeta-potential (>25 mV) with a good yield of the process (>65%). The set-up spray drying parameters allowed to obtain good yield of the process (>50%) and spherically shaped particles with the volume-weighted mean diameter (d[4,3])<6 microm in the respirable range. The set-up conditions for the preparation of the lipid dried powders did not adversely affect the structural integrity of the encapsulated pDNA. The powders kept a good transfection efficiency as compared to the fresh colloidal formulations. Lipid-based spray dried powders allowed the development of stable and viable formulations for respiratory gene delivery. In vitro dispersibility and deposition studies are in progress to determine the aerosolisation properties of the powders.
Cell-penetrating Peptide-coated Liposomes for Drug Delivery Across the Blood-Brain Barrier
Anticancer Res2019 Jan;39(1):237-243.PMID: 30591464DOI: 10.21873/anticanres.13103
Background/aim: Glioma is a deadly form of brain cancer. Doxorubicin is cytotoxic against glioma cells. However, the blood-brain barrier (BBB) limits its ability to be delivered to the brain.
Materials and methods: Liposomes (R8PLP) formed from, 1,2-Dioleoyl-3-trimethylammonium-propane chloride (DOTAP), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy-(polyethylene glycol)-2000] (PEG-DSPE), cholesterol and egg phosphatidylcholine (ePC) were modified by cell-penetrating peptide R8 conjugated with oleic acid as a novel method for delivering doxorubicin. The antitumor effect of R8PLP was evaluated by uptake, cytotoxicity and brain accumulation.
Results: The size of R8PLP was 95 nm. Doxorubicin was loaded into R8PLP by active loading with more than 95% encapsulation efficiency. Cellular uptake of R8PLP by U87-MG cells was 8.6-fold higher than that of unmodified liposomes. R8PLP reduced cell viability by 16.18% and 18.11% compared to cholesterol-ePC-liposomes and free doxorubicin, respectively, at 3.6 米M after 24 h treatment. The biodistribution of doxorubicin in the brain was significantly improved by R8PLP. The area under the concentration-time curve (AUC0.5-12 h) of R8PLP was 2.4-times higher than that of cholesterol-ePC-PEG-DSPE-liposomes.
Conclusion: These results suggest that R8-conjugated oleic acid-modified liposomes are effective delivery vehicles for glioma.
Phase behavior, DNA ordering, and size instability of cationic lipoplexes. Relevance to optimal transfection activity
J Biol Chem2001 Dec 14;276(50):47453-9.PMID: 11564736DOI: 10.1074/jbc.M105588200
Mechanisms of cationic lipid-based nucleic acid delivery are receiving increasing attention, but despite this the factors that determine high or low activity of lipoplexes are poorly understood. This study is focused on the fine structure of cationic lipid-DNA complexes (lipoplexes) and its relevance to transfection efficiency. Monocationic (N-(1-(2,3-dioleoyloxy)propyl),N,N,N-trimethylammonium chloride, N-(1-(2,3-dimyristyloxypropyl)-N,N-dimethyl-(2-hydroxyethyl)ammonium bromide) and polycationic (2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanammonium trifluoroacetate) lipid-based assemblies, with or without neutral lipid (1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine, 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine, cholesterol) were used to prepare lipoplexes of different L(+)/DNA(-) charge ratios. Circular dichroism, cryogenic-transmission electron microscopy, and static light scattering were used for lipoplex characterization, whereas expression of human growth hormone or green fluorescent protein was used to quantify transfection efficiency. All monocationic lipids in the presence of inverted hexagonal phase-promoting helper lipids (1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine, cholesterol) induced appearance of Psi(-) DNA, a chiral tertiary DNA structure. The formation of Psi(-) DNA was also dependent on cationic lipid-DNA charge ratio. On the other hand, monocationic lipids either alone or with 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine as helper lipid, or polycationic 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanammonium trifluoroacetate-based assemblies, neither of which promotes a lipid-DNA hexagonal phase, did not induce the formation of Psi(-) DNA. Parallel transfection studies reveal that the size and phase instability of the lipoplexes, and not the formation of Psi(-) DNA structure, correlate with optimal transfection.
The Impact of Lipid Types and Liposomal Formulations on Osteoblast Adiposity and Mineralization
Molecules2018 Jan 2;23(1):95.PMID: 29301300DOI: 10.3390/molecules23010095
Recent studies have demonstrated that fat accumulation in bone cells is detrimental to bone mass. Both adipocytes and osteoblasts are derived from common multipotent mesenchymal stem cells (MSCs) and hence the presence of fat may increase adipocyte proliferation, differentiation and fat accumulation while inhibiting osteoblast differentiation and bone formation. Lipids are common constituents in supramolecular vesicles (e.g., micelles or liposomes) that serve as drug delivery systems. Liposomal formulations such as Meriva? were proven to decrease joint pain and improve joint function in osteoarthritis (OA) patients. In this study, we evaluated how lipid types and liposomal formulations affect osteoblast behavior including cell viability, differentiation, mineralization and inflammation. Various liposomal formulations were prepared using different types of lipids, including phosphatidylcholine (PC), 1,2-dioleoyl-sn-glycero-3-phospho-ethanolamine (DOPE), cholesterol (Chol), 3汿[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol hydrochloride (DC-cholesterol HCl), and 1,2-Dioleoyl-3-trimethylammonium-propane chloride salt (DOTAP) to investigate the impact on osteoblast differentiation and inflammation. The results indicated that cationic lipids, DC-cholesterol and DOTAP, presented higher dose-dependent cytotoxicity and caused high level of inflammatory responses. Due to the natural properties of lipids, all the lipids can induce lipid droplet formation in osteoblasts but the level of lipid droplet accumulation was different. In comparison with cationic lipids, neutral lipids induced less adiposity, and maintained high osteoblast mineralization. Similar to previous researches, we also confirmed an inverse relationship between lipid droplet formation and osteoblast mineralization in 7F2 mouse osteoblasts. Importantly, PC containing liposomes (PC only and PC/DOTAP) suppressed IL-1汿induced gene expression of COX-2 and MMP-3 but not Chol/DOTAP liposomes or DC-Chol/DOPE liposomes. Taken together, we suggested that PC contained liposomes could provide the best liposomal formulation for the treatment of bone diseases.
Carbon dots stabilized silver-lipid nano hybrids for sensitive label free DNA detection
Biosens Bioelectron2019 May 15;133:48-54.PMID: 30909012DOI: 10.1016/j.bios.2019.03.027
Carbon dots have been extensively used for the development of fluorescent based molecular affinity sensors. However, label free DNA sensing by electrochemical method is not reported so far. Herein, we report carbon dots stabilized silver nanoparticles (CD-AgNPs) lipid nano hybrids as a sensitive and selective platform for label free electrochemical DNA sensing. The CD-AgNPs were synthesized by wet chemical method and then characterized by UV-visible, Fourier-transform Infra-red (FT-IR), dynamic light scattering (DLS) and high resolution transmission electron microscopy (HR-TEM) techniques. These CD-AgNPs were used for decorating the binary lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP) surface (named as lipid) and tethered on self-assembled monolayer of 3-mercaptopropionic acid (MPA) (MPA-lipid-CD-AgNPs). The formation of array of MPA-lipid-CD-AgNPs on Au electrode was confirmed by atomic force microscopy (AFM). Electrochemical behavior of MPA- lipid-CD-AgNPs was monitored in the presence of 1 mM potassium ferri/ferrocyanide (K3/K4 [Fe(CN)6]). The formation of layer-by-layer MPA-lipid-CD-AgNPs is indicated by increased anodic and cathodic peak (忖Ep) separation with decreased redox peak current of K3/K4 [Fe(CN)6]. Short chain DNA (30 mer oligonucleotide, representing the lung cancer) was used as a model system for label free DNA sensing. Un-hybridized (single stranded DNA), hybridized (complementary hybridized), single, double and triple base mismatched target DNA hybridized surfaces were efficiently discriminated at 1 ?M target DNA concentration at the Au/MPA-lipid-CD-AgNPs electrode by change in the charge transfer resistance from impedance technique. Further, the modified electrode was successfully used to determine target DNA in a wide linear range from 10-16 to 10-11 M. The present work open doors for the utilization of CDs in molecular affinity based electrochemical sensor design and development.